Fogle Jace L, O'Connell John P, Fernandez Erik J
Department of Chemical Engineering, University of Virginia, 102 Engineers' Way, P.O. Box 400741, Charlottesville, VA 22904-4741, USA.
J Chromatogr A. 2006 Jul 21;1121(2):209-18. doi: 10.1016/j.chroma.2006.04.015. Epub 2006 May 11.
Amide hydrogen-deuterium exchange labeling has been used to study the effects of salt and protein loading on alpha-lactalbumin (BLA) stability during hydrophobic interaction chromatography (HIC). Stability in the adsorbed phase increased dramatically with increasing loading, and unfolding was nearly undetectable close to the resin saturation capacity. We also found that a butyl surface destabilized BLA more than a phenyl surface, despite the fact that BLA was bound more strongly on the phenyl surface. These observations have important implications for HIC process design and indicate that in some cases column capacity does not have to be sacrificed to preserve protein stability.
酰胺氢-氘交换标记已被用于研究在疏水相互作用色谱（HIC）过程中盐和蛋白质负载量对α-乳白蛋白（BLA）稳定性的影响。随着负载量增加，吸附相中的稳定性显著提高，并且在接近树脂饱和容量时几乎检测不到蛋白质展开。我们还发现，尽管BLA在苯基表面上的结合更强，但丁基表面比苯基表面更能使BLA不稳定。这些观察结果对HIC工艺设计具有重要意义，并表明在某些情况下，不必为了保持蛋白质稳定性而牺牲柱容量。
